Laser Scanning Confocal Microscope Diagram

3 non confocal top and confocal bottom image of a double.

Laser scanning confocal microscope diagram.

Laser excitation wavelength is 561nm supercontinuum laser. However instead of the standard tungsten halogen or mercury arc discharge lamp one or more laser systems are used as a light source to excite fluorophores in the specimen. Its applications include research on caries response of hard and soft. Fluorescent microscopy not only makes our images look good it also allows us to gain a better understanding of cells structures and tissue.

Among various devices confocal laser scanning microscopy clsm is considered to be a valuable tool in dentistry research. Confocal microscopy can be considered a bridge between these two classical methodologies. All laser scanning confocal microscope designs are centered around a conventional upright or inverted research level optical microscope. The point to point lateral x y resolution of a spinning disk confocal microscope is essentially the same as that in laser scanning confocal or widefield fluorescence microscopy see figure 4 b.

Unlike traditional fluorescence microscopy in which the excitation wavelength is shorter than the emission wavelength two photon excitation requires simultaneous excitation by two photons with longer wavelength than the emitted light. Two photon excitation microscopy tpef or 2pef is a fluorescence imaging technique that allows imaging of living tissue up to about one millimeter in thickness. The objective is an olympus 20x water immersion na 0 54. Confocal microscopy most frequently confocal laser scanning microscopy clsm or laser confocal scanning microscopy lcsm is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out of focus light in image formation.

With confocal laser scanning microscopy clsm we can find out even more. A thick 16 micrometer section of fluorescently stained. This means that we can view visual sections of tiny structures that. The fluorescence is collected through a bandpass filter centered at 600 nm with 37 nm bandpass.

Clsm combines high resolution optical imaging with depth selectivity which allows us to do optical sectioning. The classical abbe equation describes the relationship between spatial resolution the excitation wavelength and the numerical aperture of the. Illustrated in figure 1 are a series of images that compare selected viewfields in traditional widefield and laser scanning confocal fluorescence microscopy. Diagram 5 excitation photon flux at different laser powers diagram 6 excited state saturation behavior absorb ed photons.

Confocal laser scanning microscopy principles microscopy from carl zeiss optical image formation. The pinhole aperture is 2 airy units.